Identification of an SAS (Sp1c Adjacent Site)-Like Element in the Distal 5′-Flanking Region of the Rat Lutropin Receptor Gene Essential for Cyclic Adenosine 3′,5′-Monophosphate Responsiveness

Shiyou Chen; Xuebo Liu; Deborah L Segaloff

doi:10.1210/endo.142.5.8161

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Identification of an SAS (Sp1c Adjacent Site)-Like Element in the Distal 5′-Flanking Region of the Rat Lutropin Receptor Gene Essential for Cyclic Adenosine 3′,5′-Monophosphate Responsiveness

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Identification of an SAS (Sp1c Adjacent Site)-Like Element in the Distal 5′-Flanking Region of the Rat Lutropin Receptor Gene Essential for Cyclic Adenosine 3′,5′-Monophosphate Responsiveness

Shiyou Chen, Xuebo Liu and Deborah L Segaloff

Endocrinology (Philadelphia), Vol.142(5), pp.2013-2021

05/01/2001

DOI: 10.1210/endo.142.5.8161

PMID: 11316768

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Abstract

One of the hallmarks of the differentiation of granulosa cells is the estradiol and FSH/cAMP-dependent induction of the LH receptor (LHR). Previous studies using granulosa cells isolated from diethylstilbestrol-pretreated immature rats identified a novel cAMP-responsive element termed the SAS site (Sp1c adjacent site) in the promoter region of the rat (r) LHR gene. The studies presented herein show that there is an additional distal site located at nucleotide (nt) -933/-924 that appears to interact with the same transcription factor that binds to the promoter SAS site. Similar to the SAS site, the complex formed between granulosa cell nuclear extracts and this distal site is enhanced by cAMP treatment of the granulosa cells. The core sequence required for the formation of the DNA/protein complex at this distal rLHR site was determined to be AGTGG(A)GGGG. With the exception of adenine at -928, substitution of any residue within this sequence prevented formation of this complex. The core sequence of this distal site differs from that of the proximal SAS site, which is GGGGG, and hence the distal site has been termed a SAS-like site. Reporter gene assays using constructs containing the -2,109/-1 region of the rLHR demonstrate that mutation of the distal SAS-like site abolishes the cAMP-induced transcription of the rLHR gene in rat granulosa cells, underscoring the functional significance of this site. Given the lack of sequences in the 5'-flanking region of the rLHR gene consistent with known cAMP-responsive elements, the identification of the novel SAS and SAS-like sites in the rLHR gene provides important clues toward understanding the mechanisms by which the rLHR gene is induced by FSH/cAMP.

Details

Title: Subtitle: Identification of an SAS (Sp1c Adjacent Site)-Like Element in the Distal 5′-Flanking Region of the Rat Lutropin Receptor Gene Essential for Cyclic Adenosine 3′,5′-Monophosphate Responsiveness
Creators: Shiyou Chen - Department of Physiology and Biophysics, The University of Iowa College of Medicine, Iowa City, Iowa 52242
Xuebo Liu - Department of Physiology and Biophysics, The University of Iowa College of Medicine, Iowa City, Iowa 52242
Deborah L Segaloff - Department of Physiology and Biophysics, The University of Iowa College of Medicine, Iowa City, Iowa 52242
Resource Type: Journal article
Publication Details: Endocrinology (Philadelphia), Vol.142(5), pp.2013-2021
DOI: 10.1210/endo.142.5.8161
PMID: 11316768
ISSN: 0013-7227
eISSN: 1945-7170
Language: English
Date published: 05/01/2001
Academic Unit: Molecular Physiology and Biophysics; Obstetrics and Gynecology
Record Identifier: 9984083227502771

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